摘要目的　研究血管紧张素Ⅱ对ECV304细胞中转录因子NF-κB的作用和对PDGF-B 基因表达的影响.方法　采用电泳迁移率移动分析法(EMSA)和免疫组化方法,包括共聚焦显微镜及金颗粒标记免疫电镜技术；荧光素酶报告基因与变异型激酶质粒共转染的方法及Northern 印迹法研究血管紧张素Ⅱ激活NF-κB的信号传递路径和检测了血管紧张素Ⅱ刺激前后PDGF-B mRNA的表达水平.结果　血管紧张素Ⅱ刺激后,在ECV304细胞内有NF-κB的激活及核易位过程,应用免疫荧光共聚焦显微镜,免疫电镜及Northern 印迹等方法均可观察到PDGF-B或其基因表达增高.变异型激酶质粒IKKα-KM,IKKβ-KM 及 NIK-KM 可抑制经AngⅡ刺激的转染细胞内与NF-κB启动相连的荧光素酶的表达.结论　AngⅡ可激活胞浆内NF-κB并出现核易位,激酶NIK、IKKα和IKKβ参与了此信号传递路径.血管紧张素Ⅱ刺激后PDGF-B链mRNA水平增高.

abstractsObjective　To examine the effect of angiotensin Ⅱ (Ang Ⅱ) on nuclear factor-kappa B (NF-κB) activation in human endothelial cell line ECV304 and the molecular mechanism by which Ang Ⅱ activates NF-κB. Methods　ECV304 cells were transiently cotransfected with an NF-κB/ uciferase reporter gene and inactive NF-κB-inducing kinase (NIK), IκB kinase α (IKKα), IκB kinase β (IKKβ) mutants or vectors, respectively. The effect on NF-κB was detected by using an electrophoretic mobility shift assay (EMSA) and overexpression of the mutants enabled blocking of reporter gene activation induced by Ang Ⅱ. With immunofluorescence and immuno-electronic microscope techniques, including confocal microscopy and gold particle labeled electronic microscopy, definite cytoplasmic-to-nuclear translocations of NF-κB activation were detected using subunits p50 and p65 induced by Ang Ⅱ. Results　The translocation of p50 in nuclei was highly remarkable 2 hours after Ang Ⅱ stimulation, and the activity was somewhat reduced 6 hours after stimulation to the 18th hour. Northern blot also showed PDGF-B mRNA increased by stimulation of Ang Ⅱ for 18 hours. Conclusion　Ang Ⅱ is effective in stimulating NF-κB activation through a pathway dependent on NIK, IKKα and IKKβ, and induces PDGF-B transcription in the endothelial cell line, ECV304.